17-613
ChIPAb+ p53 - ChIP Validated Antibody and Primer Set
from mouse
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About This Item
UNSPSC-Code:
12352203
eCl@ss:
32160702
NACRES:
NA.52
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Allgemeine Beschreibung
All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+ p53 set includes the p53 antibody raised against recombinant human p53, the negative control antibody (mouse normal IgG), and qPCR primers flanking an Sp1 binding site (to which p53 can be recruited1) within the human p21 (WAF1/CIP1/CDKN1A) promoter, amplifying a 105 base pair PCR product. The p53 and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of p53 associated chromatin.
The ChIPAb+ p53 set includes the p53 antibody raised against recombinant human p53, the negative control antibody (mouse normal IgG), and qPCR primers flanking an Sp1 binding site (to which p53 can be recruited1) within the human p21 (WAF1/CIP1/CDKN1A) promoter, amplifying a 105 base pair PCR product. The p53 and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of p53 associated chromatin.
Spezifität
Recognizes p53 in wild type and mutant forms of human origin.
Anwendung
Research Category
Epigenetik & nukleäre Funktionen
Epigenetik & nukleäre Funktionen
Research Sub Category
Chromatin-Biologie (ChIP)
Chromatin-Biologie (ChIP)
Chromatin Immunoprecipitation Analysis:
Sonicated chromatin prepared from untreated or actinomycin D treated (30 ng/mL, 6 hrs.) U2OS cells (3 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 1 μg of mouse Anti-p53 and the Magna ChIP® G (Cat. # 17-611) Kit. Immunoprecipitation of p53 associated DNA fragments was verified by qPCR using ChIP Primers p21 flanking the human p21 promoter that contains an Sp1 binding site (Please see figures).
Fold Increase is a ratio of normalized mean IP quantities extracted from standard curves derived from inputs of each chromatin sample. As previously published, p53 association with this region of the p21 promoter increases upon actinomycin D treatment.
Chromatin Immunoprecipitation Analysis:
Sonicated chromatin prepared from untreated or UV treated (6 hrs, 50 joules/m2.) U2OS cells (3 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 1 μg of mouse Anti-p53 and the Magna ChIP A (Cat. # 17-610) Kit.
Immunoprecipitation of p53 associated DNA fragments was verified by qPCR using ChIP Primers p21 flanking the human p21 promoter that contains an Sp1 binding site (Please see figures).
Fold Increase is a ratio of normalized mean IP quantities extracted from standard curves derived from inputs of each chromatin sample. p53 immunoprecipitable activity associated with this promoter increases with UV treatment as observed in other studies.
Sonicated chromatin prepared from untreated or actinomycin D treated (30 ng/mL, 6 hrs.) U2OS cells (3 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 1 μg of mouse Anti-p53 and the Magna ChIP® G (Cat. # 17-611) Kit. Immunoprecipitation of p53 associated DNA fragments was verified by qPCR using ChIP Primers p21 flanking the human p21 promoter that contains an Sp1 binding site (Please see figures).
Fold Increase is a ratio of normalized mean IP quantities extracted from standard curves derived from inputs of each chromatin sample. As previously published, p53 association with this region of the p21 promoter increases upon actinomycin D treatment.
Chromatin Immunoprecipitation Analysis:
Sonicated chromatin prepared from untreated or UV treated (6 hrs, 50 joules/m2.) U2OS cells (3 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 1 μg of mouse Anti-p53 and the Magna ChIP A (Cat. # 17-610) Kit.
Immunoprecipitation of p53 associated DNA fragments was verified by qPCR using ChIP Primers p21 flanking the human p21 promoter that contains an Sp1 binding site (Please see figures).
Fold Increase is a ratio of normalized mean IP quantities extracted from standard curves derived from inputs of each chromatin sample. p53 immunoprecipitable activity associated with this promoter increases with UV treatment as observed in other studies.
This ChIPAb+ p53 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Verpackung
25 assays per kit. ~1 μg per chromatin immunoprecipitation
Qualität
Routinely evaluated by chromatin immunoprecipitation and immunoblot on RIPA lysate of human Raji cells.
Zielbeschreibung
53 kDa
Physikalische Form
Protein A purified
Anti-p53 (mouse monoclonal IgG). One vial containing 25 μg of of protein A purified mouse IgG2a in 25 μL of storage buffer (PBS, pH 7.4, containing 0.05% sodium azide and 1 mg/mL BSA). Store at -20°C.
Normal Mouse IgG. One vial containing 25 μg of normal mouse IgG in 25 μL volume. Store at -20°C.
ChIP Primers p21. One vial containing 75 μL of 5 μM of each primer specific for a region of the human p21 (WAF1/CIP1/CDKN1A) promoter. Store at -20°C.
FOR: GTG GCT CTG ATT GGC TTT CTG
REV: CTG AAA ACA GGC AGC CCA AG
Normal Mouse IgG. One vial containing 25 μg of normal mouse IgG in 25 μL volume. Store at -20°C.
ChIP Primers p21. One vial containing 75 μL of 5 μM of each primer specific for a region of the human p21 (WAF1/CIP1/CDKN1A) promoter. Store at -20°C.
FOR: GTG GCT CTG ATT GGC TTT CTG
REV: CTG AAA ACA GGC AGC CCA AG
Format: Purified
Lagerung und Haltbarkeit
Stable for 1 year at -20°C from date of receipt
Hinweis zur Analyse
Control
Included negative control antibody mouse normal IgG and control primers specific for human p21 (WAF1/CIP1/CDKN1A) promoter.
Included negative control antibody mouse normal IgG and control primers specific for human p21 (WAF1/CIP1/CDKN1A) promoter.
Rechtliche Hinweise
MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Haftungsausschluss
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Lagerklassenschlüssel
10 - Combustible liquids
Analysenzertifikate (COA)
Suchen Sie nach Analysenzertifikate (COA), indem Sie die Lot-/Chargennummer des Produkts eingeben. Lot- und Chargennummern sind auf dem Produktetikett hinter den Wörtern ‘Lot’ oder ‘Batch’ (Lot oder Charge) zu finden.
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